Unlock instant, AI-driven research and patent intelligence for your innovation.

A method for identification of dragon fruit germplasm using ssr molecular markers of transcriptome sequencing

A transcriptome sequencing and molecular marker technology, applied in the field of molecular biology, can solve the problems of small proportion of pitaya genome, low marker density, inability to distinguish homozygotes and heterozygotes, etc., and achieves rich polymorphism, uniform distribution, and operation. Easy and fast effects

Active Publication Date: 2021-04-30
GUIZHOU UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the disadvantages that most of the existing molecular markers of dragon fruit are dominant markers, homozygotes and heterozygotes cannot be distinguished, and the proportion of the genome covering the dragon fruit is small, and the marker density is low, the present invention aims to provide polymorphisms with a high detection rate. , better resolution, co-dominance, stable and reliable, simple and efficient molecular marker identification method

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method for identification of dragon fruit germplasm using ssr molecular markers of transcriptome sequencing
  • A method for identification of dragon fruit germplasm using ssr molecular markers of transcriptome sequencing
  • A method for identification of dragon fruit germplasm using ssr molecular markers of transcriptome sequencing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] With EST-SSR primers in the present invention, identify 70 parts of dragon fruit germplasm, including 50 parts of red-skinned and red-fleshed type, 10 parts of red-skinned and white-fleshed type and 10 parts of red-skinned and pink-fleshed type. Taking primer pair C30929 as an example, you can Differentiate the test material.

[0032] 1. Design and synthesis of EST-SSR primers

[0033] First, pre-process the sequence obtained by sequencing to obtain high-quality EST sequences without redundancy, and use MISA software to search for SSR sites in the transcription data. The search criteria are: single nucleotide, dinucleotide, trinucleotide , four nucleotides, five nucleotides and six nucleotides, the minimum number of repetitions were 10, 6, 5, 5, 5 and 5, and then use the Primer3.0 primer batch design program for the Unigene sequence containing the SSR site Primers were designed, and the length of the SSR site sequence was between 18-24bp. Wherein, the main parameter o...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for identifying pitaya germplasm by using SSR molecular markers of transcriptome sequencing. The method designs EST-SSR marker primers shown in the sequence table and includes the following steps: S1, taking the young and tender stem tips of pitaya, It is used to extract genomic DNA; S2, SSR primers are designed based on the sequence of dragon fruit transcriptome sequencing, and the PCR amplification system is optimized; S3, PCR adopts 10 μl reaction system: 1-2 μl containing 10-50ng template DNA, forward and reverse Each primer is 0.2-0.5μl, 5μl Mix and appropriate amount of ddH 2 O; S4, the PCR reaction program is: 94°C pre-denaturation for 5 min; 94°C for 30s, 54-63°C for 30s, 72°C for 30s, 35 cycles; 72°C for 8min; 4°C for final storage; S5, PCR The amplification products are firstly screened by primers, and then by polymorphism screening, and the results are determined according to the presence or absence and size of the bands. The key technology to be solved in the present invention is the acquisition and screening of EST-SSR marker primers, and the optimization of the PCR amplification system.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to SSR primers, PCR amplification and application thereof developed based on the sequencing sequence of the dragon fruit transcriptome. Multiple pairs of SSR marker primers developed based on the transcriptome sequence are specially used for the rapid identification and genetic relationship analysis of dragon fruit germplasm from the DNA level. Background technique [0002] Dragon fruit (Hylocereus spp.) belongs to the genus Hylocereus and Seleniereus of the family Cactaceae, and is a new tropical and subtropical fruit that has been widely concerned in recent years. Dragon fruit is suitable for a wide range of planting, strong resistance to diseases and insect pests, high yield, good benefits and low market risk. Therefore, dragon fruit planting is regarded as an agricultural development project with good market prospects and economic benefits. Accurate and efficient identification a...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6858
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 文晓鹏杨仕美乔光
Owner GUIZHOU UNIV